Developmental expression of the rat muscle glycogen phosphorylase gene is not directly regulated by the four muscle regulatory factor proteins

B. Froman, Fredric A Gorin

Research output: Contribution to journalArticle

Abstract

The muscle isozyme of glycogen phosphorylase (MGP)is developmentally and neurally regulated in rat muscle. A series ot deletion derivatives were generated in a CAT reporter vector to identify elements in the 5' noncoding region of MGP required for developmental regulation. A 269 bp fragment was identified that conferred 15-fold higher levels of CAT activity in C2C12 myotubes than myoblasts. This fragment contained four E box motifs located at -167, -118, -18 and +29, a CArG-like element (CCAAAATAGC) at -109, a GC rich region at -63 and a TATA motif 27 bp 5' of the transcriptional start point. Point mutations of the E boxes had little affect on the developmental regulation of MGP. Three of the muscle regulatory factor proteins (MyoD, myogenin and MRF4) transactivated the MGP promoter by only 2-fold and one them ( myf-5) had no effect on CAT activities in cotransfection studies in the nonmuscle cell line C3H10T1/2. Deletion of the CArG-like sequence reduced the difference between MT and MB CAT activity to only 1.8-fold. This may be a potential area of importance in the developmental-specific regulation of MGP in muscle cells. Further deletion of the GC motif eliminated any measurable CAT activity and defined the minimal promoter region between -76 and +3.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number3
StatePublished - 1996

Fingerprint

Glycogen Phosphorylase
phosphorylase
glycogen
Muscle
Rats
Genes
Isoenzymes
Muscles
muscles
isozymes
rats
MyoD Protein
Proteins
genes
proteins
promoter regions
E-Box Elements
GC Rich Sequence
Myogenin
Cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

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title = "Developmental expression of the rat muscle glycogen phosphorylase gene is not directly regulated by the four muscle regulatory factor proteins",
abstract = "The muscle isozyme of glycogen phosphorylase (MGP)is developmentally and neurally regulated in rat muscle. A series ot deletion derivatives were generated in a CAT reporter vector to identify elements in the 5' noncoding region of MGP required for developmental regulation. A 269 bp fragment was identified that conferred 15-fold higher levels of CAT activity in C2C12 myotubes than myoblasts. This fragment contained four E box motifs located at -167, -118, -18 and +29, a CArG-like element (CCAAAATAGC) at -109, a GC rich region at -63 and a TATA motif 27 bp 5' of the transcriptional start point. Point mutations of the E boxes had little affect on the developmental regulation of MGP. Three of the muscle regulatory factor proteins (MyoD, myogenin and MRF4) transactivated the MGP promoter by only 2-fold and one them ( myf-5) had no effect on CAT activities in cotransfection studies in the nonmuscle cell line C3H10T1/2. Deletion of the CArG-like sequence reduced the difference between MT and MB CAT activity to only 1.8-fold. This may be a potential area of importance in the developmental-specific regulation of MGP in muscle cells. Further deletion of the GC motif eliminated any measurable CAT activity and defined the minimal promoter region between -76 and +3.",
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AB - The muscle isozyme of glycogen phosphorylase (MGP)is developmentally and neurally regulated in rat muscle. A series ot deletion derivatives were generated in a CAT reporter vector to identify elements in the 5' noncoding region of MGP required for developmental regulation. A 269 bp fragment was identified that conferred 15-fold higher levels of CAT activity in C2C12 myotubes than myoblasts. This fragment contained four E box motifs located at -167, -118, -18 and +29, a CArG-like element (CCAAAATAGC) at -109, a GC rich region at -63 and a TATA motif 27 bp 5' of the transcriptional start point. Point mutations of the E boxes had little affect on the developmental regulation of MGP. Three of the muscle regulatory factor proteins (MyoD, myogenin and MRF4) transactivated the MGP promoter by only 2-fold and one them ( myf-5) had no effect on CAT activities in cotransfection studies in the nonmuscle cell line C3H10T1/2. Deletion of the CArG-like sequence reduced the difference between MT and MB CAT activity to only 1.8-fold. This may be a potential area of importance in the developmental-specific regulation of MGP in muscle cells. Further deletion of the GC motif eliminated any measurable CAT activity and defined the minimal promoter region between -76 and +3.

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